LOCUS       sequence-319570-        9387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 RNA-dependent RNA 
            polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9387)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9387)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4133773
FEATURES             Location/Qualifiers
     source          1..9387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3382..3402
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3403..3429
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3498..3619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3509..3528)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3563..3582
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3734..3780
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3810..3946
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3991..4062
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4288..4876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4368..4387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5376..6585
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6422..6441
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6589..7617
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    7644..7692
                     /note="synthetic polyadenylation signal"
     CDS             join(7958..8026,8027..8818)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8176..8195)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8831..8967
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9119..9190
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(9195..9241)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9259..9307
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgtctgc cgacgcacag
      601 tcgttcctga atagagtttg tggtgtatcc gcggcccggc taactccatg cggaaccggg
      661 acgagcacag atgtggtgta ccgagctttt gatatctata acgacaaagt cgctggcttc
      721 gccaagtttc ttaagaccaa ctgttgcagg ttccaagaga aagacgaaga tgataatctc
      781 atagactcat actttgtggt taagcgtcac acttttagta attatcagca tgaggaaaca
      841 atttataact tgctgaagga ctgccctgcc gtagcaaaac acgacttctt taaattccgc
      901 atcgatggcg atatggtccc gcatattagc agacagcggt taaccaagta cactatggca
      961 gacctggtgt acgccctgcg acacttcgac gagggaaact gtgacacgtt aaaagaaatc
     1021 ctcgtgactt ataattgttg cgatgacgat tatttcaaca agaaagattg gtacgatttt
     1081 gtcgagaatc ccgatatact cagagtgtac gctaacctgg gggaaagggt tcgccaggca
     1141 ctattgaaga cagtccaatt ttgcgacgcg atgcggaatg ccggcattgt gggtgtcctg
     1201 acacttgata accaggacct gaatggcaac tggtatgact ttggagattt cattcagacc
     1261 acgcccgggt ccggagtacc tgtcgttgac tcttactact cacttctgat gccaatccta
     1321 accttaaccc gagctctcac agctgagagt cacgtggaca ccgacctcac aaaaccgtat
     1381 atcaagtggg atttgctgaa gtacgatttc acggaagaga ggttaaagct ctttgatcgt
     1441 tatttcaaat actgggacca gacttatcat cccaactgtg tgaattgtct ggacgacaga
     1501 tgcatacttc attgtgccaa tttcaatgta ctgtttagca cagtgttccc accaacttcg
     1561 tttggtcctc tagtgcgcaa aatctttgtg gatggggtgc ccttcgttgt ttccacaggc
     1621 tatcactttc gtgagctcgg cgtagttcac aaccaagatg tcaacctgca ttcttccagg
     1681 ttaagtttta aggaacttct ggtgtatgcc gcagaccctg cgatgcacgc tgcaagcggt
     1741 aatttgttgc tggataagcg gaccacgtgc ttcagcgtgg cggctttaac aaataacgtt
     1801 gccttccaaa ccgtcaagcc tggcaacttc aataaagatt tctacgattt cgccgtcagt
     1861 aaagggtttt ttaaggaggg atcaagcgta gaattgaaac attttttctt tgctcaggac
     1921 ggtaatgccg ctatctcgga ctacgactac taccggtata acctaccaac tatgtgcgac
     1981 attcggcagc tgttgttcgt ggtggaggtc gttgataaat actttgattg ctacgatggc
     2041 ggatgcatta acgccaatca agtcatcgtg aacaatctgg acaagagcgc cggcttcccc
     2101 tttaacaaat ggggaaaggc aaggctatat tatgactcta tgtcctatga ggatcaggac
     2161 gctctcttcg cgtataccaa gcgcaacgtt atccccacaa ttacccagat gaacctcaaa
     2221 tatgcaatca gcgcaaagaa cagagcgagg acagttgctg gggtgagtat atgttcgaca
     2281 atgactaaca ggcagtttca ccagaagctg ttgaaatcta ttgccgctac acgaggggct
     2341 accgtcgtga ttggcacctc caaattctac ggagggtggc ataatatgtt aaaaactgtg
     2401 tactccgacg tggaaaaccc tcacctgatg ggttgggatt atcctaagtg cgaccgggcg
     2461 atgccaaaca tgctgagaat tatggcgtca ttagtcctgg cgcgaaagca caccacgtgc
     2521 tgtagtctgt cacatcgctt ttacagattg gctaatgaat gcgcccaagt gcttagcgag
     2581 atggtgatgt gtgggggttc actttacgtg aagcccggag gaacgtcctc aggcgacgcc
     2641 acaactgcct acgcaaattc agtattcaac atatgtcagg ccgttacagc aaacgtgaat
     2701 gctctgctgt ctactgatgg gaacaagatc gccgataagt acgtccggaa cctgcaacac
     2761 aggctgtacg agtgtctgta tcgtaatcga gatgtagaca ccgacttcgt gaacgaattt
     2821 tacgcatact tacggaaaca tttctccatg atgatcctct ccgacgatgc agttgtgtgc
     2881 tttaattcaa cttacgcctc tcagggcttg gtggccagta tcaaaaattt taagtcggtt
     2941 ctttattatc agaataacgt gtttatgagc gaggctaagt gttggaccga gactgatctg
     3001 accaaaggcc cgcacgagtt ctgttcccag cacaccatgc tggtcaagca gggtgatgac
     3061 tatgtctatt taccataccc agatccgagt aggatcttgg gagccggttg ttttgtggat
     3121 gacattgtga agactgacgg tacactcatg attgagcgct tcgtcagttt ggcaatcgac
     3181 gcttaccccc ttaccaaaca tccaaatcag gaatatgcgg atgtgtttca cctgtacctt
     3241 caatacataa ggaaacttca tgatgaacta actggacata tgctcgacat gtattctgta
     3301 atgcttacaa atgacaatac ctcacggtac tgggaaccgg agttttatga agccatgtac
     3361 accccacata ctgtcctgca agaaaacctt tactttcagt cgtaccccta tgacgtacca
     3421 gattatgcgt aaggatccgc aggcctctgc tagcttgact gactgagata cagcgtacct
     3481 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3541 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3601 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3661 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc
     3721 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     3781 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga
     3841 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     3901 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc
     3961 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca
     4021 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg
     4081 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg
     4141 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg
     4201 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa
     4261 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4321 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4381 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4441 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4501 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4561 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4621 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4681 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4741 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4801 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4861 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4921 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4981 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5041 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5101 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac
     5161 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc
     5221 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga
     5281 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc
     5341 cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc
     5401 tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag ggcgcaggag
     5461 cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc
     5521 ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca
     5581 ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct
     5641 gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg
     5701 gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt
     5761 gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc
     5821 cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag
     5881 caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt
     5941 cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat
     6001 ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg
     6061 ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac tgactggaga
     6121 actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc
     6181 cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat
     6241 aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg
     6301 cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg
     6361 gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag
     6421 ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt
     6481 taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagacttgt
     6541 aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat gggtaaaaag
     6601 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
     6661 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
     6721 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
     6781 tatcggcact ttgcatccgc cgcgctcccg attccggaag tgcttgacat tggggagttc
     6841 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
     6901 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggcaatgga tgccatcgct
     6961 gccgccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
     7021 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
     7081 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
     7141 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
     7201 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
     7261 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
     7321 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc ccggctccgg
     7381 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
     7441 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
     7501 gtcgggcgta cacaaatcgc ccgcagaagc gccgccgtct ggaccgatgg ctgtgtagaa
     7561 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaataagct
     7621 agtatgtaag cctagtctta gataataaaa tcgctatcca tcgaagatgg atgtgtgttg
     7681 gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg
     7741 agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc
     7801 aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt
     7861 taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa
     7921 catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac atttccgtgt
     7981 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     8041 ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca tcgaactgga
     8101 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc caatgatgag
     8161 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     8221 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac cagtcacaga
     8281 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     8341 tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg agctaaccgc
     8401 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     8461 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacctt
     8521 gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt tgatagactg
     8581 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     8641 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     8701 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     8761 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaacc
     8821 gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
     8881 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
     8941 tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga ctctggctct
     9001 atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg
     9061 catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca
     9121 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
     9181 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     9241 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
     9301 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
     9361 agttcatagc ccatatatgg agttccg
//
